ABSTRACT
OBJECTIVE： To separate and purify Alhagi sparsifolia n-butanol extract monomeric compounds， and to investigate its effects on the proliferation and metastasis of human cervical cancer HeLa cells. METHODS： The n-butanol extract was separated and purified by silica gel column， Sephadex LH-20 gel column and prep-HPLC. The structures of compounds were analyzed and identified according to physicochemical properties and spectrum （mass spectrum， hydrogen spectrum， carbon spectrum） data. Using human cervical cancer HeLa cells as objects， 5-FU as positive control， MTT assay was used to detect the inhibitory rate of HeLa cells pretreated with different doses of compounds （6.25， 12.5， 25， 50， 100， 200 μg/mL）； IC50 was calculated to screen active monomers. Scratch test was used to investigate the effects of above active monomers （all 50 μg/mL） on the migration ability of HeLa cells. Kim’s formula was used to evaluate the effects of 5-FU separately combined with above active monomers [（3.125+6.25），（6.25+12.5），（12.5+25），（25+50）μg/mL]. RESULTS： Six compounds were isolated from the n-butanol extract part of A. sparsifolia and identified as butin （Ⅰ）， 3′，4′，7-trihydroxyisoflavone （Ⅱ）， p-methoxyphenylacetic acid （Ⅲ）， 4-hydroxyacetophenone （Ⅳ）， aurantiamide acetate （Ⅴ）， protocatechualdehydea （Ⅵ）. Compared with blank control group， 5-FU and each compound （5-FU：6.25-200 μg/mL， compound Ⅰ： 12.5-200 μg/mL； compound Ⅱ： 25， 50， 200 μg/mL； compound Ⅲ： 6.25， 100， 200 μg/mL； compound Ⅳ： 50， 100， 200 μg/mL； compound Ⅴ： 12.5， 25， 200 μg/mL； compound Ⅵ： 6.25-200 μg/mL） could significantly increase the cell inhibition rate. IC50 of compound Ⅰ， Ⅴ， Ⅵ were decreased significantly （P＜0.05 or P＜0.01）， and those of compound Ⅰ and Ⅵ were lower relatively. The migration distance of cells in 5-FU and compound Ⅰ and Ⅵgroups were decreased significantly， compared with blank control group （P＜0.05 or P＜0.01）. 5-FU separately combined with compound Ⅰ and Ⅵ showed additive and enhanced inhibitory effects on the proliferation of HeLa cells （synergistic index＞0.9）. CONCLUSIONS： Compounds Ⅰ-Ⅵ are isolated from Alhagi for the first time. Butin and protocatechualdehydea are active monomers of its n-butanol extract part. Above two monomers can inhibit the proliferation and migration of human cervical cancer Hela cells， with strong inhibitory effect in vitro， and stronger inhibitory effect combined with 5-FU than any compound alone.
ABSTRACT
Objective:To compare and analyze the chemical constituents of volatile oils extracted from the different parts ( flow-ers,leaves and roots) of Stelleropsis tianschanica by chromatography-mass spectrometry (GC-MS). Methods:The volatile oil was ex-tracted by diethyl ether-Soxhlet extraction method and analyzed by GC-MS with a capillary gas chromatographic column. The relative contents of the volatile compounds were calculated by chromatographic peak area normalization method.Results: Totally 179 volatile constituents in the different parts of Stelleropsis tianschanica were identified. Among them,81 compounds were identified in leaves,and the relative content accounted for 82.77% of the total volatile compounds;108 compounds were identified in flowers,and the relative content accounted for 82.85% of the total volatile compounds;112 compounds were identified in roots, and the relative content ac-counted for 85.98% of the total volatile compounds. Totally 33 compounds existed in all the three parts,and the content accounted for 39.24% of the total volatile components in leaves,35.86% in flowers and 48.89% in roots. The relative content of(Z,Z)-9,12-oc-tadecadienoic acid in leaves,flowers and roots of S. tianschanica was the highest,which accounted for 11.12%,9.8% and 22.49%, respectively. Conclusion:The different parts of S. tianschanica have similar volatile components, while the specific substances and the contents are different.